High-Level Expression of Immunogenic Recombinant Plasmo-dium vivax Merozoite Surface Protein (Pvmsp-1 42 kDa) in pGEX 6P1 Vector

Seyyed Tabaei, Seyyed Javad and Haghighi, Ali and Kazemi, Bahram and Fallah Omrani, Vahid and Fallahi, Shirzad and MIRAHMADI, Hadi (2015) High-Level Expression of Immunogenic Recombinant Plasmo-dium vivax Merozoite Surface Protein (Pvmsp-1 42 kDa) in pGEX 6P1 Vector. Iran J Public Health, 44 (1). pp. 89-99.

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Abstract

Background: Detection of Plasmodium vivax specific antibodies with serological tests could be a valuable tool for epi-demiological researches. Whereas P. vivax cannot be simply obtained in vitro, serological tests using total or semi-purified antigens are infrequently used. Given this restriction, the present study investigated whether recombinant P. vivax merozoite surface protein 1 (PvMSP-1 42 kDa) could be useful in detection of antibodies from the serums of a P. vivax infected person using serological tests. Methods: Parasite DNA was extracted from blood sample of an Iranian P. vivax-infected patient. The region of PvMSP-142 kDa was amplified by PCR then cloned into pTZ57R/T vector and sequenced. The insert was sub cloned into pGEX 6P1 expression vector. Afterwards, it was transformed into E. coli BL21 and cultured massively. Sub clon-ing of gene was confirmed by PCR and enzyme digestion and sequencing finally. Production of recombinant protein was confirmed by SDS-PAGE. Western blot was performed by human sera to appraisal binding ability to the IgG antibodies of P. vivax infected patients . Recombinant protein was purified and estimated by Bradford assay. Results: The specialty values of the Western blot determined with 10 sera from naturally infected individuals, 10 sera from healthy individuals and 7 sera from individuals with other infectious diseases. Conclusion: For the Iranian population, using a Western blot assay for MSP-142 recombinant protein can be used as the foundation for promotion of serological assay for the detection of P. vivax malaria such as ELISA. Keywords: Plasmodium vivax, Recombinant PvMSP-1 42 kDa, Expression vector, Iran

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